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Establishment of effective gene disruption tools for gene functional analysis in basidiomycetous fungi.

Research Project

Project/Area Number 22780149
Research Category

Grant-in-Aid for Young Scientists (B)

Allocation TypeSingle-year Grants
Research Field Forest science
Research Institution公益財団法人岩手生物工学研究センター (2011)
Iwate Biotechnology Research Center (2010)

Principal Investigator

SAKAMOTO Yuichi  公益財団法人岩手生物工学研究センター, 生物資源研究部, 主任研究員 (80390889)

Project Period (FY) 2010 – 2011
Project Status Completed (Fiscal Year 2011)
Budget Amount *help
¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2011: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
Fiscal Year 2010: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Keywords森林生産 / 育種 / 遺伝子破壊 / シイタケ / 担子菌類 / 食用きのこ
Research Abstract

To establish effective gene disruption tool for edible basidiomyceteous fungi(mushrooms), gene disruption vector system and resistant marker gene recycling system were constructed in Lentinula edodes. First, a gene disruption vector system was constructed by using DelsGate method, and then the loxP sequences were inserted the vector construction system. Unfortunately, no gene disrupted mutant was obtained yet. The pcc1 gene that is a negative regulator in monokaryotic strain, and the ku80 that is involved in non-homologous DNA joint were cloned in L. edodes. If the gene could disrupted, the strain can form fruiting body without mating, and the strain have high efficiency for homologous recombination. To test Cre-loxP system in basidiomyceteous fungi, faster than L. edodes, loxP sequences were inserted gene disruption cassette for C. cinerea. Marker recycling system will be test in C. cinerea in the near future.

Report

(3 results)
  • 2011 Annual Research Report   Final Research Report ( PDF )
  • 2010 Annual Research Report
  • Research Products

    (27 results)

All 2012 2011 2010

All Journal Article (11 results) (of which Peer Reviewed: 10 results) Presentation (11 results) Book (2 results) Patent(Industrial Property Rights) (3 results)

  • [Journal Article] Characterization of beta-N-acetylhexosaminidase(LeHex20A), a member of glycoside hydrolase family 20, from Lentinula edodes(shiitake mushroom)2012

    • Author(s)
      Konno N., Takahashi H., Nakajima M., Takeda T., Sakamoto Y.
    • Journal Title

      AMB express

      Volume: 2 Issue: 1 Pages: 29-29

    • DOI

      10.1186/2191-0855-2-29

    • Related Report
      2011 Final Research Report
    • Peer Reviewed
  • [Journal Article] Characterization of beta-N-acetylhexosaminidase (LeHex20A), a member of glycoside hydrolase family 20, from Lentinula edodes (shiitake mushroom)2012

    • Author(s)
      Naotake Konno, Hideyuki Takahashi, Masahiro Nakajima, Takumi Takeda, Yuichi Sakamoto
    • Journal Title

      AMB express

      Volume: (印刷中)

    • Related Report
      2011 Annual Research Report
    • Peer Reviewed
  • [Journal Article] An endo-β-1, 6-glucanase involved in Lentinula edodes fruiting body autolysis2011

    • Author(s)
      Naotake Konno and Yuichi Sakamoto
    • Journal Title

      Applied Microbiology and Biotechnology

      Volume: 91(5) Issue: 5 Pages: 1365-1373

    • DOI

      10.1007/s00253-011-3295-2

    • Related Report
      2011 Annual Research Report 2011 Final Research Report
    • Peer Reviewed
  • [Journal Article] Endo-β-1, 3-glucanase, GLU1, from Lentinula edodes fruiting body belongs to a new glycoside hydrolase family2011

    • Author(s)
      Sakamoto Y., Nakade K., Konno N.
    • Journal Title

      Applied and Environmental Microbiology

      Volume: 77(23) Issue: 23 Pages: 8350-8354

    • DOI

      10.1128/aem.05581-11

    • Related Report
      2011 Annual Research Report 2011 Final Research Report
    • Peer Reviewed
  • [Journal Article] シイタケ収穫後の子実体老化に関与する酵素の研究2011

    • Author(s)
      坂本裕一
    • Journal Title

      日本きのこ学会誌

      Volume: 19(2) Pages: 73-78

    • NAID

      110008711281

    • URL

      http://ci.nii.ac.jp/naid/110008711281

    • Related Report
      2011 Annual Research Report 2011 Final Research Report
    • Peer Reviewed
  • [Journal Article] Gene silencing of the Lentinula edodes lcc1 gene by expression of a homologous inverted repeat sequence2011

    • Author(s)
      Nakade K., Watanabe H., Sakamoto Y., Sato T.
    • Journal Title

      Microbiological Research

      Volume: 166 Issue: 6 Pages: 484-493

    • DOI

      10.1016/j.micres.2010.09.004

    • Related Report
      2011 Final Research Report
    • Peer Reviewed
  • [Journal Article] An endo-β-1, 6-glucanase involved in Lentinula edodes fiuiting body autolysis2011

    • Author(s)
      Konno N., Sakamoto Y.
    • Journal Title

      Applied Microbiology and Biotechnology

      Volume: (印刷中)

    • Related Report
      2010 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Gene silencing of the Lentinula edodes lcc1 gene by expression of a homologous inverted repeat sequence.2011

    • Author(s)
      Nakade K., Watanabe H., Sakamoto Y., Sato T.
    • Journal Title

      Microbiological Research

      Volume: (印刷中)

    • Related Report
      2010 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Characterization of an extracellular laccase, PbLac1, purified from Polyporus brumalis2010

    • Author(s)
      Nakade K., Nakagawa Y., Yano A., Sato T., Sakamoto Y.
    • Journal Title

      Fungal Biology

      Volume: 114 Issue: 8 Pages: 609-618

    • DOI

      10.1016/j.funbio.2010.05.002

    • Related Report
      2011 Final Research Report 2010 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Protein expression during Flammulina velutipes fruiting body formation2010

    • Author(s)
      Yuichi Sakamoto
    • Journal Title

      Mycoscience

      Volume: 51 Issue: 3 Pages: 163-169

    • DOI

      10.1007/s10267-010-0032-0

    • NAID

      10027043455

    • Related Report
      2011 Final Research Report
    • Peer Reviewed
  • [Journal Article] Sakamoto Y.(2010) Protein expression during Flammulina velutipes fruiting body formation2010

    • Author(s)
      Sakamoto, Y.
    • Journal Title

      Mycoscience

      Volume: 51 Pages: 163-169

    • Related Report
      2010 Annual Research Report
  • [Presentation] 環境刺激に応答するきのこの子実体発生2012

    • Author(s)
      坂本裕一
    • Organizer
      静電気学会バイオプロセス・プラズマ研究会
    • Place of Presentation
      岩手大学
    • Related Report
      2011 Annual Research Report 2011 Final Research Report
  • [Presentation] 分泌型ラッカーゼ(Lcc1)RNAi抑制株におけるシイタケ細胞壁構造の変化2011

    • Author(s)
      中出啓子、坂本裕一
    • Organizer
      糸状菌分子生物学コンファレンス
    • Year and Date
      2011-11-16
    • Related Report
      2011 Annual Research Report
  • [Presentation] Transcriptome analysis of the autolysis after harvesting of the Lentinula edodes fruiting body2011

    • Author(s)
      坂本裕一
    • Organizer
      Asian Mycological Congress
    • Place of Presentation
      仁川、韓国
    • Year and Date
      2011-08-10
    • Related Report
      2011 Annual Research Report
  • [Presentation] Transcriptome analysis of the autolysis after harvesting of the Lentinula edodes fruiting body2011

    • Author(s)
      Yuichi Sakamoto
    • Organizer
      Asian Mycological Congress
    • Place of Presentation
      韓国、仁川
    • Related Report
      2011 Final Research Report
  • [Presentation] 分泌型ラッカーゼ(Lcc1) RNAi抑制株におけるシイタケ細胞壁構造の変化2011

    • Author(s)
      中出啓子、坂本裕一
    • Organizer
      糸状菌分子生物学コンファレンス
    • Place of Presentation
      東京大学
    • Related Report
      2011 Final Research Report
  • [Presentation] Enzymes involved in fruiting body senescence in L.edodes.2010

    • Author(s)
      Sakamoto, Y., Nakade, K., Konno, N.
    • Organizer
      The 6th Meeting of East Asia for Mushroom Science
    • Place of Presentation
      韓国、慶州
    • Year and Date
      2010-11-14
    • Related Report
      2010 Annual Research Report
  • [Presentation] シイタケにおける形態形成と細胞壁分解酵素の発現挙動2010

    • Author(s)
      坂本裕一
    • Organizer
      日本きのこ学会2010年度大会
    • Place of Presentation
      東京大学(招待講演)
    • Year and Date
      2010-09-16
    • Related Report
      2010 Annual Research Report
  • [Presentation] Characterization of biological function of laccases in Lentinula edodes2010

    • Author(s)
      Sakamoto, Y., Nakade, K
    • Organizer
      International Mycological Congress 9th
    • Related Report
      2011 Final Research Report
  • [Presentation] シイタケにおける形態形成と細胞壁分解酵素の発現挙動2010

    • Author(s)
      坂本裕一
    • Organizer
      日本きのこ学会2010年度大会
    • Place of Presentation
      東京大学
    • Related Report
      2011 Final Research Report
  • [Presentation] Enzymes involved in fruiting body senescence in L. edodes2010

    • Author(s)
      Sakamoto, Y., Nakade, K., Konno, N.
    • Organizer
      The 6th Meeting of East Asia for Mushroom Science
    • Place of Presentation
      韓国、慶州
    • Related Report
      2011 Final Research Report
  • [Presentation] Characterization of biological function of laccases in Lentinula edodes.2010

    • Author(s)
      Sakamoto, Y., Nakade, K.
    • Organizer
      International Mycological Congress 9th
    • Place of Presentation
      スコットランド、エジンバラ
    • Related Report
      2010 Annual Research Report
  • [Book] Food Quality2012

    • Author(s)
      Yuichi Sakamoto, Keiko Nakade, Naotake Konno, Toshitsugu Sato
    • Publisher
      Intech
    • Related Report
      2011 Final Research Report
  • [Book] タイトル:Senescence of the Lentinula edodes fruiting body after harvesting書名:Food Quality2012

    • Author(s)
      Yuichi Sakamoto, Keiko Nakade, Naotake Konno, Toshitsugu Sato
    • Total Pages
      28
    • Publisher
      Intech
    • Related Report
      2011 Annual Research Report
  • [Patent(Industrial Property Rights)] β-N-アセチルヘキソサミニダーゼ2012

    • Inventor(s)
      金野尚武、坂本裕一
    • Industrial Property Rights Holder
      岩手県
    • Filing Date
      2012-02-14
    • Related Report
      2011 Final Research Report
  • [Patent(Industrial Property Rights)] β-N-アセチルヘキソサミニダーゼ2012

    • Inventor(s)
      金野尚武、坂本裕一
    • Industrial Property Rights Holder
      岩手県
    • Industrial Property Number
      2012-009380
    • Filing Date
      2012-02-14
    • Related Report
      2011 Annual Research Report
  • [Patent(Industrial Property Rights)] L-DOPAを蓄積する植物細胞及びその利用2011

    • Inventor(s)
      中塚貴司、高橋秀行、山田恵理、坂本裕一、関口拓史、佐々木伸大、小関良宏、西原昌宏
    • Industrial Property Rights Holder
      岩手県
    • Filing Date
      2011-03-22
    • Related Report
      2011 Final Research Report

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Published: 2010-08-23   Modified: 2016-04-21  

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